*Target (protein/gene name): NADH-dependent Enoyl–Acyl carrier protein reductase *NCBI Gene # or RefSeq#: Gene ID:886523 (ncbi) http://www.uniprot.org/uniprot/P0A5Y6 *Protein ID (NP or XP #) or Wolbachia#: NP_216000.1 *Organism (including strain): Mycobacterium tuberculosis Etiologic Risk Group: Risk Group 3 Bacterial Agents Including Rickettsia *Background/Disease Information (sort of like the Intro to your Mini Research Write up): The bacteria Mycobacterium tuberculosis tends to mainly attack the human lungs and cause tuberculosis or TB. If it is not treated, TB can prove to be fatal. The bacteria passes through the hair and can infect nearby people who breath in the bacteria. Mycobacterium tuberculosis can cause latent TB infection or TB disease. The enzyme enoyl-acyl carrier protein reductase in M. tuberculosis is an important an functional protein for type II fatty acid synthesis. Unfortunately, some strains of TB are resistant to Isoniazid and other multiple drugs that try to inhibit this protein to stop infection. Essentiality of this protein: Rv1484 has essentiality data Gene/Ortholog: mtu1508 (OG4_13747); Phenotype: non-essential; Source study: nmpdr Gene/Ortholog: eco1251 (OG4_13747); Phenotype: undefined; Source study: blattner Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: gerdes Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: keio Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: shigen
Assay was not found on Sigma webstite Structure Available (PDB ): 1ENY Current Inhibitors: 2 natural inhibitors in strain A Expression Information (has it been expressed in bacterial cells):
yes; it has been expressed in E. coli Purification Method: The protein can be purified using recombinant His-tagged InhA from E. coli BL2 (DE3) by Ni-NTA affinity column chromatography and gel filtration. Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): MTGLLDGKRI LVSGIITDSS IAFHIARVAQ EQGAQLVLTG FDRLRLIQRI TDRLPAKAPL LELDVQNEEH LASLAGRVTE AIGAGNKLDG VVHSIGFMPQ TGMGINPFFD APYADVSKGI HISAYSYASM AKALLPIMNP GGSIVGMDFD PSRAMPAYNW MTVAKSALES VNRFVAREAG KYGVRSNLVA AGPIRTLAMS AIVGGALGEE AGAQIQLLEE GWDQRAPIGW NMKDATPVAK TVCALLSDWL PATTGDIIYA DGGAHTQLL *length of your protein in Amino Acids: 268 Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 28527.8 Molar Extinction coefficient of your protein at 280 nm wavelength: 30940 TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. *CDS Gene Sequence (paste as text only):
*GC% Content for gene: *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized): Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol) -- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):
*NCBI Gene # or RefSeq#: Gene ID:886523 (ncbi)
http://www.uniprot.org/uniprot/P0A5Y6
*Protein ID (NP or XP #) or Wolbachia#: NP_216000.1
*Organism (including strain): Mycobacterium tuberculosis
Etiologic Risk Group: Risk Group 3 Bacterial Agents Including Rickettsia
*Background/Disease Information (sort of like the Intro to your Mini Research Write up): The bacteria Mycobacterium tuberculosis tends to mainly attack the human lungs and cause tuberculosis or TB. If it is not treated, TB can prove to be fatal. The bacteria passes through the hair and can infect nearby people who breath in the bacteria. Mycobacterium tuberculosis can cause latent TB infection or TB disease. The enzyme enoyl-acyl carrier protein reductase in M. tuberculosis is an important an functional protein for type II fatty acid synthesis. Unfortunately, some strains of TB are resistant to Isoniazid and other multiple drugs that try to inhibit this protein to stop infection.
Essentiality of this protein:
Rv1484 has essentiality data
Gene/Ortholog: mtu1508 (OG4_13747); Phenotype: non-essential; Source study: nmpdr
Gene/Ortholog: eco1251 (OG4_13747); Phenotype: undefined; Source study: blattner
Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: gerdes
Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: keio
Gene/Ortholog: eco1251 (OG4_13747); Phenotype: essential; Source study: shigen
Complex of proteins: yes
Druggable Target: yes; 0.7 index
*EC#: 1.3.1.9
Link to BRENDA EC# page:
http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.3.1.9&Suchword=&organism%5B%5D=Mycobacterium+tuberculosis&show_tm=0
Enzyme Assay information (spectrophotometric, coupled assay, reagents):
Assay of Mycobacterium smegmatis:
- http://www.brenda-enzymes.info/literature/lit.php4?e=1.3.1.44&r=390808
- http://www.brenda-enzymes.info/literature/lit.php4?e=1.3.1.44&r=390807
Reagents of Mycobacterium tuberculosis:- http://www.brenda-enzymes.info/literature/lit.php4?e=1.3.1.44&r=657270
- http://www.brenda-enzymes.info/literature/lit.php4?e=1.3.1.44&r=390808
Assay was not found on Sigma webstiteStructure Available (PDB ): 1ENY
Current Inhibitors: 2 natural inhibitors in strain A
Expression Information (has it been expressed in bacterial cells):
yes; it has been expressed in E. coli
Purification Method: The protein can be purified using recombinant His-tagged InhA from E. coli BL2 (DE3) by Ni-NTA affinity column chromatography and gel filtration.
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MTGLLDGKRI LVSGIITDSS IAFHIARVAQ EQGAQLVLTG FDRLRLIQRI TDRLPAKAPL
LELDVQNEEH LASLAGRVTE AIGAGNKLDG VVHSIGFMPQ TGMGINPFFD APYADVSKGI
HISAYSYASM AKALLPIMNP GGSIVGMDFD PSRAMPAYNW MTVAKSALES VNRFVAREAG
KYGVRSNLVA AGPIRTLAMS AIVGGALGEE AGAQIQLLEE GWDQRAPIGW NMKDATPVAK
TVCALLSDWL PATTGDIIYA DGGAHTQLL
*length of your protein in Amino Acids: 268
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 28527.8
Molar Extinction coefficient of your protein at 280 nm wavelength: 30940
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
Primer design results for 'tail' primers (this is just 2 sequences):